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Microorganisms
Volume 9
Issue 6
10.3390/microorganisms9061170
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Article

Scanning Electron Microscope: A New Potential Tool to Replace Gram Staining for Microbe Identification in Blood Cultures

by
Gabriel Haddad
1,2,
Sara Bellali
1,
Tatsuki Takakura
3,
Anthony Fontanini
1,
Yusuke Ominami
4,
Jacques Bou Khalil
1,* and
Didier Raoult
1,2,*
1
Institut Hospitalo-Universitaire Méditerranée Infection 19-21 Boulevard Jean Moulin, 13005 Marseille, France
2
Aix-Marseille Université, IRD, APHM, MEPHI, 27 Boulevard Jean Moulin, 13005 Marseille, France
3
Hitachi High-Tech Corporation, Analytical & Medical Solution Business Group, 882 Ichige, Hitachinaka-shi 312-8504, Ibaraki, Japan
4
Hitachi High-Tech Corporation, Nanotechnology Solutions Business Group, Toranomon Hills Business Tower, 1-17-1 Toranomon, Minato-ku, Tokyo 105-6409, Japan
*
Authors to whom correspondence should be addressed.
Microorganisms 2021, 9(6), 1170; https://doi.org/10.3390/microorganisms9061170
Submission received: 16 May 2021 / Revised: 25 May 2021 / Accepted: 26 May 2021 / Published: 28 May 2021
(This article belongs to the Section Medical Microbiology)
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Abstract

Blood culture is currently the most commonly used method for diagnosing sepsis and bloodstream infections. However, the long turn-around-time to achieve microbe identification remains a major concern for clinical microbiology laboratories. Gram staining for preliminary identification remains the gold standard. We developed a new rapid strategy using a tabletop scanning electron microscope (SEM) and compared its performance with Gram staining for the detection of micro-organisms and preliminary identification directly from blood cultures. We first optimised the sample preparation for twelve samples simultaneously, saving time on imaging. In this work, SEM proved its ability to identify bacteria and yeasts in morphotypes up to the genus level in some cases. We blindly tested 1075 blood cultures and compared our results to the Gram staining preliminary identification, with MALDI-TOF/MS as a reference. This method presents major advantages such as a fast microbe identification, within an hour of the blood culture being detected positive, low preparation costs, and data traceability. This SEM identification strategy can be developed into an automated assay from the sample preparation, micrograph acquisition, and identification process. This strategy could revolutionise urgent microbiological diagnosis of infectious diseases.
Keywords: scanning electron microscopy; gram; MALDI-TOF; blood culture; rapid test; microbe identification scanning electron microscopy; gram; MALDI-TOF; blood culture; rapid test; microbe identification

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MDPI and ACS Style

Haddad, G.; Bellali, S.; Takakura, T.; Fontanini, A.; Ominami, Y.; Bou Khalil, J.; Raoult, D. Scanning Electron Microscope: A New Potential Tool to Replace Gram Staining for Microbe Identification in Blood Cultures. Microorganisms 2021, 9, 1170. https://doi.org/10.3390/microorganisms9061170

AMA Style

Haddad G, Bellali S, Takakura T, Fontanini A, Ominami Y, Bou Khalil J, Raoult D. Scanning Electron Microscope: A New Potential Tool to Replace Gram Staining for Microbe Identification in Blood Cultures. Microorganisms. 2021; 9(6):1170. https://doi.org/10.3390/microorganisms9061170

Chicago/Turabian Style

Haddad, Gabriel, Sara Bellali, Tatsuki Takakura, Anthony Fontanini, Yusuke Ominami, Jacques Bou Khalil, and Didier Raoult. 2021. "Scanning Electron Microscope: A New Potential Tool to Replace Gram Staining for Microbe Identification in Blood Cultures" Microorganisms 9, no. 6: 1170. https://doi.org/10.3390/microorganisms9061170

APA Style

Haddad, G., Bellali, S., Takakura, T., Fontanini, A., Ominami, Y., Bou Khalil, J., & Raoult, D. (2021). Scanning Electron Microscope: A New Potential Tool to Replace Gram Staining for Microbe Identification in Blood Cultures. Microorganisms, 9(6), 1170. https://doi.org/10.3390/microorganisms9061170

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